Title: Prokaryotic Expression and Identification of Murine Lipocalin 2
Abstract: Objective To express murine lipocalin 2 (lcn2) and study its biological activity. Methods Extract total RNA from murine macrophage RAW264.7 strain for amplification of lcn2 gene by RT-PCR. Insert the amplified lcn2 gene into prokaryotic expression vector pET-32a (+), and transform the constructed recombinant plasmid pET-32a (+)-lcn2 to E. coli BL21 (DE3) plysS for expression under induction of IPTG. The expressed product was identified by His-tag in-gel staining, purified by Ni2+-NTA affinity chromatography and determined for biological activity. Results Both nucleotide sequencing and restriction analysis proved that recombinant plasmid pET-32a(+)-lcn2 was constructed correctly. SDS-PAGE showed that expressed recombinant lcn2 protein, with a relative molecular mass of about 21 000, mainly existed in a form of inclusion body and contained 35% of total somatic protein. The expressed product reached a protein concentration of 1. 0 g / L after purification and showed a certain inhibitory effect on both E.coli and β-streptococcus. Conclusion Recombinant lcn2 protein was successfully expressed in prokaryotic cells and showed a certain bacleriostatic effect.
Publication Year: 2008
Publication Date: 2008-01-01
Language: en
Type: article
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